肿瘤细胞裂解物致敏的树突状细胞对结肠癌LoVo细胞的杀伤作用

目的 探讨肿瘤细胞裂解物致敏的树突状细胞(DC)作为佐剂对结肠癌LoVo细胞系的杀伤作用.方法 反复冻融法裂解培养的结肠癌LoVo细胞,提取细胞抗原并致敏DC,然后将后者与自身T淋巴细胞共同培养,观察其对自身T淋巴细胞增殖的影响以及活化的T淋巴细胞对LoVo细胞的杀伤作用.结果 结肠癌LoVo细胞裂解物致敏的DC能明显促进T淋巴细胞的增殖,后者对LoVo细胞的杀伤作用明显增强.结论 肿瘤细胞裂解物提取方法简单,易于临床实施,其作为细胞抗原致敏DC制备的肿瘤疫苗能有效活化并产生肿瘤特异性细胞毒T淋巴细胞,将有很好的临床应用前景.     

胎儿来源的树突状细胞体外诱导抗前列腺癌效应的实验研究

目的研究胎儿来源树突状细胞(DC)体外诱导抗前列腺癌的特异性细胞免疫效果。方法从胎儿骨髓、肝脏获得单个核细胞,经重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)、重组人白细胞介素-4(rhIL-4)和重组人肿瘤坏死因子-α(rhTNF-α)诱导产生DC。50%~70%硫酸铵饱和沉淀法获取前列腺癌细胞DU145含热休克蛋白(HSP)成分的细胞溶解物,以该抗原负载DC,激活胎脾细胞产生肿瘤特异性细胞毒性T淋巴细胞(CTL)。MTT法检测CTL对DU145、PC3和EJ细胞的杀伤效应。结果胎儿骨髓、肝脏可诱导出功能成熟的DC,高表达CD1 a、CD86、HLA-DR和CD83。负载DU145抗原的DC可诱导产生CD8+CTL。CD8+T细胞表型由转化前的(14.09±2.46)%变为转化后的(62.76±2.64)%。对DU145细胞有明显细胞毒作用,显著高于对EJ细胞杀伤效应(P<0.01)。结论含HSP成分的肿瘤细胞溶解物负载胎儿来源DC,体外可诱导出特异性抗肿瘤免疫应答。     

负载冻融抗原的树突状细胞诱导特异性抗膀胱癌作用的研究

目的研究负载膀胱癌冻融抗原的树突状细胞（DC）诱导的对膀胱癌细胞的特异性杀伤效应。方法反复冻融法获得BIU-87细胞抗原;人单个核细胞在含重组人GM—CSF、IL-4和TNF-a的RPMI1640培养基中体外诱导出人DC并负载肿瘤抗原;9d后成熟DC与自体T细胞共孵育诱导膀胱癌抗原特异性CTLs; 用MTT法检测其对BIU-87体外杀伤效应;用ELISA法检测DC分泌IL-12的能力。结果负载膀胱癌抗原的DC诱导的特异性CTLs可明显杀伤BIU-87细胞,在效靶比为40：1时杀伤率为（65．5±6．4）％,显著高于各对照组（P〈0．01）;负载抗原DCs较未负载抗原DCs有更强的分泌IL-12能力（P〈0．05）,而且不同时期的DC分泌的量不同。结论负载膀胱癌抗原的DC在体外可诱导出高效而特异的抗膀胱癌效应,提示以DC为中心的肿瘤生物治疗有望提高膀胱癌综合治疗水平。     

膀胱癌患者外周血树突状细胞的体外培养扩增和鉴定

目的研究膀胱癌患者外周血树突状细胞(DC)的体外培养扩增和鉴定。方法应用Ficoll-Hypaque离心获得界面细胞,贴壁培养2h,获得单个核细胞,体外以重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF,50ng/ml)+重组人白细胞介素4(rhlL-4,10ng/ml)+人肿瘤坏死因子-α(hTNF-α,50ng/ml)诱导培养。倒置相差显微镜及扫描电镜下观察DC生长情况；流式细胞仪检测DC表型；MTT法检测DC活化的T细胞对肿瘤细胞BIU87的杀伤率。结果第6天体外培养的DC由贴壁状态变为悬浮毛刺状细胞,第8天为形态不规则的毛刺状,为典型DC形态。流式细胞仪检测表明,成熟DC高水平表达CD_(1a)、CD_(83)、CD_(86)及HLA-DR等。被DC激活的T细胞对膀胱癌细胞株BIU87的杀伤率为(48.8±3.7)%,未经DC激活的T细胞的杀伤率为(25.7±1.5)%,2组比较差异有统计学意义(P＜0.01)。结论膀胱癌患者外周血经rhGM-CSF+rhIL-4+hTNF-α体外诱导培养能诱导出DC。     

TNF-α增强树突状细胞诱导的抗胰腺癌免疫应答的实验研究

目的探讨体外扩增树突状细胞（DC）的方法，体外研究DC介导的细胞毒性T淋巴细胞（CTL）对小鼠胰腺癌的特异性杀伤作用。方法联合应用重组鼠源性粒细胞-巨噬细胞集落刺激因子（GM—CSF）和IL-4体外诱导骨髓源性DC，与肿瘤细胞溶解物共培养制备DC疫苗；观察DC在负载抗原后体外诱导的主动特异性CTL对胰腺癌细胞的杀伤作用。结果用GM—CSF和IL-4成功地在体外扩增了骨髓源性DC。TNF—α可诱导骨髓源性DC成熟，使其表达的CD54、主要组织相溶性复合物-Ⅱ、CD86等表面分子明显升高，并增强自分泌IL-12和抗原的递呈能力，与对照组比较，差异有统计学意义（P〈0.05）。未负载肿瘤细胞抗原的DC所冲击的T淋巴细胞不能有效地识别特定的靶细胞并产生杀伤作用，DC通过捕获并递呈坏死肿瘤细胞抗原才能诱发显著的主动特异性CTL，体外对胰腺癌细胞的杀伤效果非常显著（P〈0.01）。结论TNF—α可诱导DC的成熟，使DC的抗原递呈和自分泌IL-12的能力显著提高，DC递呈抗原后能诱发显著的主动特异性CTL。     

负载膀胱癌酸洗抗原肽对树突状细胞分泌IL-12的影响及意义

目的：研究负载膀胱癌酸洗抗原肽对树突状细胞（DCs）分泌IL-12的影响和意义.方法：用弱酸洗脱法获得膀胱癌细胞株BIU-87肿瘤抗原肽;联合应用重组人GM-CSF、IL-4和TNF-α体外诱导健康志愿者DCs并负载膀胱癌肿瘤抗原肽;DC诱导膀胱癌抗原特异性CTLs;用MTT法检测其对BIU-87体外杀伤效应;用ELISA法检测第3、6、9天培养细胞的上清液,评价DCs分泌IL-12 p70的能力.结果：联合应用重组人GMCSF、IL-4和TNF-α可在体外诱导出DCs;负载膀胱癌抗原肽的DC诱导的特异性CTLs可杀伤高达（78.5±7.0）%的BIU-87细胞,显著高于各对照组（P〈0.01）;经ELISA方法检测不同时期的DCs分泌IL-12的量不同,而且负载抗原肽DCs较未负载抗原DCs有更强的分泌能力（P〈0.05）.结论：IL-12 p70在DCs刺激特异性CTLs的过程中发挥重要作用,其分泌量受DCs的成熟状态及某些刺激信号的影响,膀胱癌酸洗抗原肽是其刺激信号之一.     

IFNγ修饰的DC对T细胞增值及杀伤肿瘤细胞作用影响的研究

目的:探讨结肠癌细胞抗原致敏的IFNγ修饰的树突状细胞（DC）对T淋巴细胞增殖和分化的影响，以及活化T细胞对结肠癌LoVo细胞的杀伤作用。方法:构建携带人IFNγ基因的重组复制缺陷型腺病毒载体（Ad-IFNγ），用其感染人外周血单个核细胞来源的DC;以流式细胞仪检测DC表型的变化。用反复冻融裂解法提取的结肠癌LoVo细胞抗原致敏DC，将其与自身T淋巴细胞共同培养，观察T细胞增殖和分化的情况以及活化T细胞对LoVo细胞的杀伤作用。结果:Ad-IFNγ转染后24 h内几乎不影响DC的吞噬功能，DC自身表达的IFNγ在一定程度上能促进DC的成熟; IFNγ修饰的DC经LoVo细胞抗原致敏后能明显促进T淋巴细胞的增殖并使其向Th1细胞分化，Th1细胞对结肠癌LoVo有明显的特异性杀伤效应。结论:IFNγ修饰的DC能增强促进T淋巴细胞的增殖，诱导细胞免疫，增强T细胞的细胞毒作用，从而有效地杀伤肿瘤细胞。     

肾癌树突状细胞疫苗的体外活性研究

目的 探讨负载人肾癌细胞抗原肽制备树突状细胞(DC)疫苗体外杀伤肾癌细胞的作用.方法 利用细胞膜酸洗脱法获得人肾透明细胞癌细胞株786-0细胞表面抗原肽.外周血单个核细胞在体外经人粒细胞一巨噬细胞集落刺激因子,白细胞介素4和脂多糖诱导获得成熟的DC,并负载分离到的抗原肽制备DC疫苗.利用疫苗体外诱导出特异性细胞毒性T淋巴细胞(CTL)作为实验组.同时设置4个对照组,对照组1用未负载抗原肽的DC和单个核细胞混合培养,对照组2用单个核细胞进行培养,对照组3用抗原肽与单个核细胞混合培养,对照组4用未负载抗原肽的DC单独培养.4个对照组也加入同实验组相同的各种因子进行培养.51Cr释放法检测特异性CTL的杀伤活性.结果 疫苗诱导的特异性CTL对肾癌细胞的杀伤活性为(31.93±5.05)%,与对照组(5.88±2.26)%、(8.03±6.70)%、(9.70±2.09)%、(9.35±3.58)%相比差异有统计学意义(P<0.05).结论 负载抗原肽的DC疫苗体外试验有高效的抗肾癌细胞活性.     

胃癌肿瘤裂解物修饰的树突状细胞体外诱导抗原特异性细胞毒T淋巴细胞

目的　利用树突状细胞呈递肿瘤抗原的特性提高细胞毒T淋巴细胞 (CTLs)对胃癌细胞的杀伤活性。方法　胃细胞癌患者外周血来源的有核细胞体外经GM -CSF和IL -4诱导产生树突状细胞 ,负载肿瘤裂解物后诱导自体CTLs产生。用细胞毒试验检测CTLs杀伤活性 ,和用ELISA测定细胞因子的分泌。结果　胃癌患者自体来源的DC裂解物能诱导产生的CTLs对自体胃癌细胞具有高杀伤率 ,可达 83 % ;致敏的DC组中IL -12与TNF -α的浓度 ( 1161± 2 3 9pg/ml,10 44± 3 12 pg/ml)显著高于未致敏的DC组 ( P <0 .0 5 )。结论　DC能呈递胃癌裂解物 ,诱导产生抗原特异性CTLs。     

负载肾癌肿瘤裂解物的树突状细胞诱导产生抗原特异性细胞毒T淋巴细胞

目的　利用树突状细胞呈递肿瘤抗原的特性提高细胞毒T淋巴细胞 (CTLs)对肾癌细胞的杀伤活性。　方法　肾细胞癌患者骨髓来源的有核细胞体外经GM CSF和IL 4诱导产生树突状细胞 ,负载肿瘤裂解物后诱导自体CTLs产生。用细胞毒试验和ELISA测定CTLs杀伤活性和细胞因子的分泌。　结果　肾癌患者自体来源的DC Tuly能 (1)增加CTLs增殖 ,16d时使T细胞增殖达 4 3倍 ;(2 )上调CTLs中CD3 和CD8 T细胞群 ;(3)诱导产生的CTLs对自体肾癌细胞具有高杀伤率 ,显著高于异体肾癌和异种肿瘤细胞 (P <0 .0 5 ) ;(4)上调TNF α分泌。　结论　DCs能呈递Tuly抗原。诱导产生抗原特异性CTLs,提示DC Tuly具有为肾癌患者制作疫苗和进行特异性CTLs过继免疫治疗的临床应用前景     

树突状细胞疫苗对人化荷人膀胱癌SCID鼠循环微转移转归的影响

目的将人外周血淋巴细胞（PBL）以腹腔转输方法构建具备人免疫环境SCID鼠模型,探讨树突状细胞（Dc）疫苗对荷瘤SCID鼠血循环膀胱癌细胞微转移转归的影响。方珐体外侵袭法从膀胱癌细胞株T24中筛选具有高转移能力亚群（T24。）;从人外周血中分离PBL,经腹腔注射小鼠（4× 10^7／鼠）,1d后,于鼠右后肢内侧皮下接种3× 10^6个T24-3细胞,2、4、5、6周分别检测鼠体内人免疫状态（包括血清IgG水平和CD3＋、CD4＋、CD8＋T细胞数量）及外周血细胞角蛋白20（CK20）mRNA表达;用含10％人AB型血清的RPMI-1640培养基,在rhGM—CSF和rhIL-4存在下,37℃、5％CO2环境下体外诱导培养人外周血单个核细胞（PBMC）来源DC。用冻融法获得的T24-3细胞裂解上清液致敏DC;并干预荷人瘤细胞的SCID鼠自发血循环肿瘤细胞微转移,观察大体情况、外周血CK20mRNA表达及瘤体MMP-7mRNA表达。结果SCID鼠于接种T24-3肿瘤细胞后5周进行DC疫苗干预。DC疫苗治疗组小鼠外周血中CK20mRNA表达均阴性,无远处转移发生;PBS处理组小鼠外周血中有2例CK20mRNA表达阳性,3例发生远处转移;DC治疗组小鼠外周血中有1例CK20mRNA表达阳性,1例发生远处转移。DC疫苗治疗组小鼠瘤组织中MMP-7mRNA表达最低,较PBS处理和DC治疗组差异有显著性意义（P〈O．01）。结论DC疫苗可以有效控制荷人膀胱癌SCID鼠血循环肿瘤细胞微转移发生,进而延缓或阻止肿瘤转移。     

Presentation of prostate tumor antigens by dendritic cells stimulates T-cell proliferation and cytotoxicity

Dendritic cells (DCs) are “professional” antigen-presenting cells capable of stimulating T-cell proliferation and cytotoxicity when loaded with and presenting specific antigens, including tumor antigens. We demonstrated the stimulation of an autologous cytotoxic T-cell response elicited by DC loaded with autologous tumor cell lysate derived from primary prostate tumor. A candidate tumor antigen is prostate-specific membrane antigen (PSMA), which is overexpressed in prostate cancer patients. We identified a HLA-A2 motif in PSMA, isolated patient DC, loaded peptide into DC, and stimulated autologous T cells to proliferate. The ability to use DC for presentation of either tumor or peptide antigen in an HLA-restricted fashion in order to stimulate T-cell proliferation and cytotoxicity demonstrates the potential of this technology for development of a prostate cancer vaccine. © 1996 Wiley-Liss, Inc.     

乳腺癌细胞抗原负载的自体树突状细胞体内诱导特异性T细胞应答

目的 以乳腺癌细胞抗原体外负载自体树突状细胞(DCs)对24例乳腺癌患者进行自身免疫,探讨其体内诱导特异性T细胞免疫应答的能力.方法 新鲜癌组织制备成热休克凋亡细胞抗原负载外周单核细胞来源DC,分别在采血后第1、2、4、6周于患者腹股沟淋巴结富集区进行皮内注射,细胞剂量为4×10+～6×106/次.结果 DC免疫治疗后患者血清抗瘤免疫因子水平明显上升:白细胞介素(IL)-2治疗前为(33.8±7.2)ng/L,治疗后为(68.5±12.4)ng/L;IL-12治疗前为(48.5±10.9)ng/L,治疗后为(118.2±31.5)ng/L;肿瘤坏死因子(TNF)-α治疗前为(18.7±5.3)ng/L,治疗后为(78.3±11.5)ng/L;干扰素(IFN)-γ治疗前为(20.5±6.3)ng/L,治疗后为(92.6±14.9)ng/L,治疗前后比较差异有统计学意义(P<0.01);DTH试验阳性率为7/10;4/10例IFN-γ+CD8+T细胞频率较治疗前明显增加.随访发现:除1例患者在治疗后3个月内疾病进展(PD),其余患者病情稳定无复发与转移症状(SD).结论 以乳腺癌细胞为抗原负载自体DC免疫患者,能够有效提高患者非特异免疫水平,激发体内特异性T细胞免疫应答,是一种安全、副反应较小、有效的乳腺癌辅助治疗手段.     

超抗原诱导细胞毒性T淋巴细胞抗膀胱肿瘤效应的研究

研究超抗原金黄色葡萄球菌肠毒素Ａ对淋巴细胞激活,增殖的作用及对Ｅ－Ｊ膀胱肿瘤细胞的抑癌效应。方法：分离健康供血者的Ｔ淋巴细胞,加ＳＥＡ培养其活化增殖,用ＭＴＴ显色法测定细胞毒性Ｔ淋巴细胞的细胞增殖动力学及对Ｅ－Ｊ膀胱细胞体外肿瘤的作用。     

树突状细胞功能异常对结直肠癌肝转移的影响

目的 探讨树突状细胞(DC)功能异常对结直肠癌肝转移的影响及其意义.方法 收集2008年1月至2010年6月健康成年人30例、结直肠癌术前、术后无肝转移者30例和术后肝转移30例患者外周血,分离单核细胞后分别加入GM-CSF(1000 U/ml)、IL-4(1000 U/ml)和'TNF-α(1000 U/ml)诱导分化成熟DC,通过加入HT29大肠癌细胞抗原处理(100 μg/ml)或不处理,DC∶T细胞为1∶10,培养7 d,用ELISA方法测定培养液中IFN-γ和IL-10水平,酶标仪测定CCK8和LDH光密度(OD)值,间接测定T细胞增殖和杀伤能力.结果 结直肠癌术后肝转移患者外周血DC在无肿瘤抗原刺激时与健康人T细胞混合培养液IL-10水平显著低于结直肠癌术前患者和健康人[(11.9±1.3)pg/ml比(29.6±9.7)pg/ml、(23.4±8.0)pg/ml,F=4.475,P＜0.05].结直肠癌术后肝转移患者外周血DC经肿瘤抗原刺激与健康人T细胞混合培养液IFN-γ水平均显著高于结直肠癌术后和健康人[(34 ± 9)pg/ml比(26±12)pg/ml、(24 ±6) pg/ml,F=5.206,P＜0.05].结直肠癌术后肝转移患者DC经HT29肠癌细胞抗原刺激后诱导健康人T淋巴细胞杀伤能力显著高于结直肠癌术前患者和健康成年人[(30.6 ±8.6)pg/ml比(12.1 ± 2.4)pg/ml、(14.9 ±1.7)pg/ml,F=4.147,P＜0.05].结论 结直肠癌患者外周血DC在肿瘤抗原刺激下诱导T淋巴细胞产生IL-10,导致肿瘤免疫逃逸,发生肝转移;外源肿瘤抗原刺激能够提高T淋巴细胞杀伤能力.

Abstract：

Objective To observe the clinical significance and effect of dysfunction of dendritic cell( DC) in colorectal cancer with liver metastasis.Methods Peripheral blood were respectively collected from healthy adult donors (30 cases), preoperative and postoperative coloreatal cancer patients without liver metastasis (30 cases) , and 30 postoperative coloreatal cancer patients with liver metastasis from Jan 2008 to Jun 2010.Peripheral blood mononuclear cells were separated, GM-CSF( 1000 U/ml) , IL-4( 1000 U/ml) and TNF-α ( 1000 U /ml) were added into cell culture fluid to induce the mononuclear cells for mature dendritic cells.There were two subgroups, and in antigen processing subgroup the lysate of HT29 colonic carcinoma cells (100 μg/ml) were added into the cell culture fluid.The T lymphocytes from healthy adults were added into two subgroups by ratio of 1∶10 ( DCs∶ T cells) , cocultured for 7 days.The level of INF-γ and IL-10 in cell culture fluid was assayed with ELISA method.The optical density (OD) of CCK8 ans LDH was assayed with ELIASA to indirectly measure the reproductive activity and the killing efficacy of T lymphocytes.Results The IL-10 level in cocultured fluid of peripheral blood DCs in postoperative colorectal carcinoma patients with liver metastasis and T lymphocytes of healthy adults was significantly higher than that of preoperative patients of colorectal carcinoma and health adults without tumor antigenic stimulation(11.9±1.3) pg/ml vs.(29.6±9.7) pg/ml, (23.4±8.0) pg/ml, F =4.475, P ＜0.05).The IFN-γ level in cocultured fluid of peripheral blood DCs in postoperative colorectal carcinoma patients with liver metastasis and T lymphocytes of healthy adults was significantly higher than that of postoperative patients of colorectal carcinoma and healthy adults with or without tumor antigenic stimulation ( 34 ± 9) pg/ml vs.(26 ± 12 ) pg/ml, (24 ± 6) pg/ml, F = 5.206, P ＜ 0.05).The killing activity of healthy adults T lymphocytes induced by HT29 colonic carcinoma cells in postoperative colorectal carcinoma patients with liver metastasis was significantly higher than that of preoperative patients of colorectal carcinoma and healthy adults (30.6 ±8.6) pg/ml vs.(12.1 ±2.4) pg/ml, (14.9 ± 1.7) pg/ml, F =4.147, P ＜ 0.05).Conclusions T lymphocytes produce IL-10 when indued by DCs from patients with colorectal carcinoma under stimulation of tumor antigen leading to tumor immune escape and liver metastasis.The killing activity of T lymphocytes can be enhanced when stimulated by exogenous tmuor antigen.     

超抗原SEA联合树突状细胞诱导特异性抗膀胱肿瘤研究

目的 观察超抗原金黄色葡萄球菌肠毒素A(SEA)联合树突状细胞(Dc)诱导CTL对膀胱肿瘤高效特异性的免疫杀伤作用和对肿瘤局部免疫提呈功能的改善情况.方法 用GM-CSF和IL-4联合刺激诱导外周血单个核细胞分化为DC.将Dc与同源淋巴细胞(DC-L组)、超抗原SEA淋巴细胞(DC-SEA-L组)共培养,用FCS分析DC供刺激分子表型,酶联免疫吸附测定法检测细胞因子IL-2,MTT法测定激活的CTL对膀胱癌E-J细胞的体外杀伤效应.结果 在体外,DCSEA-L对膀胱癌细胞具有相对特异的杀伤作用,SEA-L和Dc-L则无明显特异性.结论 SEA和Dc联合应用可诱导产生高效并具有相对特异性的抗膀胱癌效应,并改善膀胱癌组织局部的DC抗原提呈功能.     

Adriamycin-mediated potentiation of cytotoxicity against freshly isolated bladder cancer cells by autologous non-activated peripheral blood lymphocytes and tumor infiltrating lymphocytes

PURPOSE: Previous studies indicated that cancer patients lack functional anti-tumor cytotoxic lymphocytes. However, anti-tumor cytotoxic lymphocytes may coexist with immunoresistant tumor cells. We reasoned that anti-tumor cytotoxic activity of lymphocytes may be revealed if the tumor cells are sensitized to killing. It has been reported that adriamycin (ADR) exhibits various immunomodulating activities. In the present study, we investigate the effect of ADR on the susceptibility of freshly isolated bladder cancer cells to lysis by autologous non-activated peripheral blood lymphocytes (PBL) and tumor infiltrating lymphocytes (TIL). MATERIALS AND METHODS: Cytotoxicity was determined by a 1-day microculture tetrazolium dye assay. RESULTS: Treatment of ADR-resistant fresh bladder cancer cells with ADR at 0.1 microg./ml. or more for 3 hours or more enhanced their susceptibility to lysis by autologous PBL. This ADR-induced enhancement of susceptibility of fresh bladder cancer cells to lysis by PBL was also observed when lymphokine activated killer cells, purified natural killer cells and T lymphocytes were used as effector cells. Furthermore, while cytotoxicity of freshly derived TIL against autologous bladder cancer cells was minimal, significant cytotoxicity was observed with ADR-treated bladder cancer cells. The ADR analogs, epirubicin and pirarubicin, also enhanced the susceptibility of bladder cancer cells to lysis by autologous PBL. Treatment of bladder cancer cells with ADR had no effect on the expression of MHC class I on the cancer cells or the frequency of bladder cancer target cell conjugates to autologous PBL. Treatment of bladder cancer cells with ADR augmented their sensitivity to anti-Fas monoclonal antibody and tumor necrosis factor-a. Pretreatment of effector cells with ADR had no effect on their cytotoxic function. CONCLUSIONS: These findings demonstrate that PBL and TIL in patients with bladder cancer exhibit anti-tumor cytotoxic function, but their function is not manifested due to development or acquisition of tumor cell resistance to killing. However, the resistance of bladder cancer cells to killing by cytotoxic lymphocytes is overcome if cancer cells are sensitized by subtoxic concentrations of ADR. These findings suggest that treatment of bladder cancer patients with low doses of ADR may sensitize the cancer cells to killing by autologous circulating and tumor infiltrating lymphocytes and may be a novel immunotherapeutic modality for the treatment of drug-resistant and/or immune-resistant bladder cancer.     

树突状细胞瘤苗联合吉西他滨对肝癌干细胞的杀伤效应

目的:探讨负载CD133+肝癌细胞抗原的树突状细胞(DC)联合吉西他滨(GEM)在体外对肝癌干细胞的杀伤效应。方法:取对数生长期的人肝癌细胞系Huh-7以CD133作为分子标志进行流式分选,得到肝癌干细胞。将人外周血单个核细胞(PBMC)在体外诱导分化为树突状细胞(DC)。DC负载Huh-7细胞和CD133+细胞抗原后与T细胞共育得到特异性细胞毒性T细胞,将CD133+细胞作为靶细胞进行细胞毒性试验。实验组按处理因素分为:GEM组,CD133+-CTL组,GEM+CD133+-CTL组,Huh7-CTL组和GEM+Huh7-CTL组。CCK-8法检测杀伤率,然后比较各组间差异。结果:对CD133+细胞的杀伤效应以GEM+CD133+-CTL组最强,与其他各组比较差异均有统计学意义(P0.05)。单独就DC瘤苗杀伤率,CD133+-CTL组高于Huh7-CTL组(P0.05)。结论:CD133+肝癌干细胞裂解产物致敏的DC瘤苗联合化疗药物可以有效杀伤肝癌干细胞,进而可能降低肝癌术后和肝癌肝移植后的转移和复发率。